Eisuke Enoki

Cell adhesion molecule 1 is a new osteoblastic cell adhesion molecule and a diagnostic marker for osteosarcoma

AIMS An immunohistochemical screen for mouse embryos showed that cell adhesion molecule 1 (CADM1), which is an immunoglobulin superfamily member, was expressed in developing bones. Here, we determined the cell types expressing CADM1 and examined its usefulness in the differential diagnosis of osteosarcoma. MAIN METHODS Serial sections of murine developing mandibles were stained with anti-CADM1 antibody, by a coloring substrate reactive to alkaline phosphatase (ALP), a broad osteoblastic marker for preosteoblasts to osteoblasts, and by in situ hybridization for osteopontin (OPN), a marker for mature osteoblasts. CADM1 immunohistochemistry was also performed on human remodeling bones, osteosarcomas and other soft tissue tumors. KEY FINDINGS CADM1 immunohistochemistry for the mandible revealed that morphologically identifiable osteoblasts expressed CADM1 on their plasma membranes, but neither osteocytes nor bone lining cells did. At the mandibular margin, not only OPN-positive cells but also OPN-negative, ALP-positive cells were CADM1-positive, whereas inside the mandible, OPN-positive cells were often CADM1-negative. Clear membranous staining was detected in the majority of osteosarcomas (46/57), whereas only 13% (6/46) of the other soft tissue tumors were CADM1-positive (P<0.001). SIGNIFICANCE These results indicated that CADM1 was a novel osteoblastic adhesion molecule that is expressed transiently during osteoblastic maturation, and a useful diagnostic marker for osteosarcoma cells.

Congenital pneumonia with sepsis caused by intrauterine infection of Ureaplasma parvum in a term newborn: a first case report.

We present an autopsy case of intrauterine pneumonia in a term newborn in whom Ureaplasma parvum was confirmed by PCR examinations, including a novel diagnostic tool for detecting pathogens that caused neonatal infections using multiplex PCR. This is the first report of U. parvum being implicated in the pathogenesis of congenital pneumonia with sepsis in a term newborn.

Increased Ectodomain Shedding of Cell Adhesion Molecule 1 from Pancreatic Islets in Type 2 Diabetic Pancreata: Correlation with Hemoglobin A1c Levels

Pulmonary emphysema and type 2 diabetes mellitus (T2DM), both caused by lifestyle factors, frequently concur. Respectively, the diseases affect lung alveolar and pancreatic islet cells, which express cell adhesion molecule 1 (CADM1), an immunoglobulin superfamily member. Protease-mediated ectodomain shedding of full-length CADM1 produces C-terminal fragments (CTFs) with proapoptotic activity. In emphysematous lungs, the CADM1 shedding rate and thus the level of CTFs in alveolar cells increase. In this study, CADM1 expression in islet cells was examined by western blotting. Protein was extracted from formalin-fixed, paraffin-embedded sections of pancreata isolated from patients with T2DM (n = 12) or from patients without pancreatic disease (n = 8) at autopsy. After adjusting for the number of islet cells present in the adjacent section, we found that full-length CADM1 decreased in T2DM islets, while ectodomain shedding increased. Hemoglobin A1c levels, measured when patients were alive, correlated inversely with full-length CADM1 levels (P = 0.041) and positively with ectodomain shedding rates (P = 0.001). In immunofluorescence images of T2DM islet cells, CADM1 was detected in the cytoplasm, but not on the cell membrane. Consistently, when MIN6-m9 mouse beta cells were treated with phorbol ester and trypsin to induce shedding, CADM1 immunostaining was diffuse in the cytoplasm. When a form of CTFs was exogenously expressed in MIN6-m9 cells, it localized diffusely in the cytoplasm and increased the number of apoptotic cells. These results suggest that increased CADM1 ectodomain shedding contributes to blood glucose dysregulation in T2DM by decreasing full-length CADM1 and producing CTFs that accumulate in the cytoplasm and promote apoptosis of beta cells. Thus, this study has identified a molecular alteration shared by pulmonary emphysema and T2DM.

Chlamydospores of Rhizopus microsporus var. rhizopodiformis in Tissue of Pulmonary Mucormycosis

Hyphae are usually the only fungal elements found in tissue of mucormycosis, and other fungal elements are quite rarely encountered. We found chlamydospores in bronchial lumina in autopsied tissue of pulmonary mucormycosis of a diabetic patient. Chlamydospores are thick-walled, asexually produced spores arising from the modification of a hyphal segment. This is the first histologic demonstration of chlamydospores in mucormycosis in which the causative fungus is culturally identified to species level. Rhizopus microsporus var. rhizopodiformis was isolated from the present autopsied pulmonary tissue. A literature review of human infection by this fungus found 27 cases with histopathologic evidence.

Coinfection of Aspergillus and Cryptococcus in post-tuberculosis pulmonary cavity.

To the Editor: Aspergillus and Cryptococcus often infect human lungs. However, coexistence of these fungi is extremely rare in any infected site, including the lungs. We report a case of coinfection with Aspergillus and Cryptococcus in the posttuberculosis pulmonary cavity of an otherwise healthy patient. A 74-year-old man who had a history of pulmonary tuberculosis 15 years ago was admitted to a regional hospital for chest computed tomography (CT) examination because of dry cough. His routine chest radiography of recent years only detected small calcification of bilateral upper lobes. However, CT disclosed a subpleural pulmonary cavity, 28 mm in longer diameter, of the left lower lobe. The cavity wall was smooth and its thickness was regular without surrounding infiltrate and consolidation. It was considered to be old inflammatory changes related to previous tuberculosis. Thereafter, he was followed by CT every 4–6 months for 33 months without particular changes. He did not come to the regional hospital for 1 year afterwards, and CT 1 year later demonstrated irregular thickening of the cavity wall (Fig. 1a). Positron emission tomography (PET)/CT showed slight F-fluoro-2deoxyglucose uptake (SUVmax: 1.9) in the lesion. These results allowed an estimation of the probability of malignancy. Video-assisted thoracic surgery was performed to remove the cavitary lesion. Intraoperative frozen section examination revealed capsulated yeast-like fungi, consistent with Cryptococcus, and no malignant cells, therefore, no further surgery was performed. Clinical condition and laboratory data indicated that the patient was in an immunocompetent state throughout the disease course. Serum fungal antigen, fungal antibody, and b-D-glucan were not measured because fungal infection was not considered in the differential diagnosis of the pulmonary disease. The patient’s postoperative course was not eventful. Oral itraconazole therapy has been continued since the day following the operation. Grossly, the removed cavitary lesion had a subpleural location and adhered to the parietal pleura. Sectioning demonstrated a cavitary lesion, 30 mm in diameter, with a 20 mm wide lumen and 5 mm thick wall, composed of firm grayishwhite tissue with anthracosis (Fig. 1b). The cavity contained pus-like creamy material that was submitted to microbiological examination. The culture of the material grew Aspergillus sp. and Cryptococcus neoformans. Microscopically, the cavity wall was composed of hyalinized fibrous tissue with focal calcification (Fig. 1c). The inner four fifths of the cavity wall was necrotic with many small ghost cells and nuclear debris, both of which seemed to have originated from necrotic inflammatory cells. Many neutrophils infiltrated mostly in the outer layer of the cavity wall. Matted septate hyphae with dichotomous branching, consistent with an Aspergillus fungal ball, were found with necrotic material in the cavity (Fig. 1d). Yeastlike cells with capsular halo, consistent with Cryptococcus, and septate dichotomously branching hyphae coexisted in the cavity and inner layer of the cavity wall (Fig. 2a). Mucicarmin and alcian blue stained the capsules, while the wall of yeastlike cells was stained by periodic acid–Schiff (PAS), Grocott’s methenamine silver and Fontana–Masson stains (Fig. 2b). There was no granuloma and histiocytic infiltration phagocytizing yeast-like cells. Only neutrophils infiltrated close to the hyphae. Immunohistochemical analysis was kindly performed by Dr Masao Hotchi, Shinshu University, Japan, on paraffin sections of the cavitary lesion following the method of Fukuzawa et al. The septate hyphae were positive for Aspergillus antibody, while the capsulated yeast-like cells were positive for Cryptococcus antibody (Fig. 2c, d). The results of microbiology, histology and immunohistochemistry confirmed coinfection of the two fungi. Combined infection of Aspergillus and Cryptococcus in the same lesion is extremely rare, and we have found only three reports describing coexistence of these fungi in the literature; all of which showed lung cavities. In the first case, entangled hyphae, indicative of fungal balls, were seen within the cavitated cryptococcoma, with no hyphal invasion to surrounding pulmonary parenchyma. In the second case, aspergilloma in cavitated cryptococcoma finally changed to invasive aspergillosis with bone invasion. In the third case, invasive aspergillosis and cryptococcosis were seen in recently appeared pulmonary cavities. In all three reports with the infection of the two fungi, Cryptococcus played a major role in pulmonary cavitation. Cavitation is usually the result of central necrosis of cryptococcoma. The present case represents a rare scenario of Cryptococcus infection in an old tuberculous cavity with coexistence of Aspergillus. In conclusion, coinfection of Aspergillus and Cryptococcus occurred in the post-tuberculosis pulmonary cavity. The infection of these fungi should be considered when interpreting changes during follow-up of an old tuberculous cavity. bs_bs_banner

Malignant Transformation of Hepatocellular Adenoma

The patient was a 20-year-old male in whom a hepatic hypervascular mass accompanied by intratumoral hemorrhage was detected on examination for epigastric pain. Based on the enlargement of the mass and diagnostic imaging, hepatocellular adenoma (HCA) was suspected and hepatectomy was performed. The lesion was diagnosed as malignant transformation of β-catenin-activated HCA. There are only few reports of cases with malignant transformation of HCA in Japan; it is necessary to accumulate cases to investigate it.

18f-fdg Pet/ct Features of Chronic Sclerosing Sialadenitis Presenting as Localized Igg4-related Disease

Chronic sclerosing sialadenitis is a benign inflammatory condition that most commonly affects the submandibular gland in elderly individuals. It is currently known to belong to the spectrum of IgG4-related systemic diseases, which is reflected by systemic involvement on F-FDG PET/CT images. We presented a case of a 73-year-old man with histologically proven IgG4-related chronic sclerosing sialadenitis, unilateral localized form, on whole-body F-FDG PET/CT images that mimicked submandibular carcinoma with lymph node metastasis.

Needle-Tract Seeding on the Proximal Gastric Wall After EUS-Guided Fine-Needle Aspiration of a Pancreatic Mass

Needle-Tract Seeding on the Proximal Gastric Wall After EUS-Guided Fine-Needle Aspiration of a Pancreatic Mass

Research Misconduct in Japan and How It Is Covered by the Media

　Cases of research misconduct (fabrication, falsification, and plagiarism) have been increasing worldwide, including in Japan. In particular, since 2006, many cases of research misconduct have been reported in Japan, and these cases have also been covered by the media. The 2014 case of the withdrawal of articles on STAP cells followed a rare course in which research misconduct became a full-blown social phenomenon. In recent years, even the University of Tokyo has experienced reported cases of research misconduct. In this report, I would like to introduce some representative cases of research misconduct in the field of life sciences over the past decade. These examples include studies conducted at Osaka University Graduate School of Medicine (2006), Osaka University Graduate School of Frontier Bioscience (2006), Ryukyu University School of Medicine (2010), Toho University School of Medicine (2012), The University of Tokyo Institute of Molecular and Cellular Biosciences (2013), and several cases outside of Japan. I will discuss what researchers should do to reduce the incidence of research misconduct. In addition, I will discuss how these cases were covered by the media, because the publics impression of research misconduct is formed by media coverage.

Induction of Complete Remission by Azacitidine in a Patient with Myelodysplastic Syndrome-Associated Inflammatory Bowel Disease

Myelodysplastic syndrome [MDS] is a clonal disorder of bone marrow [BM] cells, caused by acquired chromosomal abnormalities and gene mutations. Pro-inflammatory antigen-presenting cells [APCs] originating from BM cells bearing chromosomal abnormalities and gene mutations can cause immune-mediated disorders including inflammatory bowel disease [IBD]. Here, we report the first case with MDS-associated IBD that was successfully treated with the DNA methyltransferase inhibitor, azacitidine [AZA]. A 75-year-old man with a 5-year history of MDS was admitted for examination of diarrhoea and high fever. Blood examination revealed pancytopenia and a marked elevation of C-reactive protein. Colonoscopy revealed multiple round ulcers from the terminal ileum to the sigmoid colon. Pathological examination of the endoscopic biopsy specimens showed destruction of crypt architecture and infiltration of CD3+ T cells and CD68+ macrophages. Surprisingly, administration of AZA, which has been approved for the treatment of high-risk MDS, improved the symptoms, and the multiple round ulcers disappeared. AZA treatment markedly decreased the expressions of tumour necrosis factor-α, interleukin-12 (IL-12)/23p40 and IL-17 in colonic biopsy samples, as assessed by quantitative reverse transcription polymerase chain reaction. In contrast, AZA treatment did not change the expression of forkhead box P3, a master regulator of regulatory T cells. These data suggest that AZA treatment led to complete remission in MDS-associated IBD through suppression of pro-inflammatory cytokine responses.